Objective: To identify the core symptoms and bridge symptoms in the network structure among adolescents bullying victimization, anxiety and depressive symptoms, and to explore the interrelationships among these three variables, so as to provide a basis for the precise prevention of bullying behaviors and the improvement of adolescents psychological health conditions. Methods: From October to November 2023, a stratified cluster random sampling method was employed to select 4 759 middle and high school students from four cities in Guangxi: Beihai, Guigang, Hechi, and Laibin. The Chinese version of the Olweus Bully/Victim Questionnaire (OBVQ) was used to assess bullying victimization among adolescents, while the Generalized Anxiety Disorder-7 (GAD-7) and the Patient Health Questionnaire-9 (PHQ-9) were utilized to assess anxiety and depressive symptoms, respectively. The Bootnet package (version 1.6) in R software (version 4.4.1) was used to construct a network structure and analyze the associations between bullying victimization, anxiety symptoms and depressive symptoms among adolescents. Results: The reporting rates of depressive symptoms, anxiety symptoms, and bullying victimization among adolescents in Guangxi were 24.67%, 16.33%, and 16.58%, respectively. Network analysis revealed that the node with the highest expected influence was "property being taken or damaged", with a standardized expected influence (EI) index of 2.09. The top two nodes in terms of bridge expected influence were "thoughts of self harm or suicide" and "irritability", with standardized bridge expected influence (BEI) indices of 2.15 and 1.27, respectively. The stronger associations were observed between the node "property being taken or damaged" and the nodes "physical attacks such as hitting, kicking, pushing, or shoving" and "ridiculed due to accent", with edge weights of 0.43 and 0.35, respectively. Conclusion Targeting preventive measures against the symptoms with the highest expected influence and bridge expected influence in the network of bullying victimization, anxiety and depressive symptoms among adolescents may be an effective approach to reduce the negative impact of bullying victimization, anxiety and depressive symptoms on adolescents.

Based on the interleukin-17(IL-17) signaling pathway, this study explores the effect and mechanism of Bufei Decoction on Klebsiella pneumoniae pneumonia in rats. SD rats were randomly divided into the control group, model group, Bufei Decoction low-dose group(6.68 g·kg~(-1)·d~(-1)), Bufei Decoction high-dose group(13.36 g·kg~(-1)·d~(-1)), and dexamethasone group(1.04 mg·kg~(-1)·d~(-1)), with 10 rats in each group. A pneumonia model was established by tracheal drip injection of K. pneumoniae. After successful model establishment, the improvement in lung tissue damage was observed following drug administration. Core targets and signaling pathways were screened using transcriptomics techniques. Real-time fluorescence quantitative polymerase chain reaction was used to detect the mRNA expression of core targets interleukin-6(IL-6), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and chemokine CXC ligand 6(CXCL6). Western blot was used to assess key proteins in the IL-17 signaling pathway, including interleukin-17A(IL-17A), nuclear transcription factor-κB activator 1(Act1), tumor necrosis factor receptor-associated factor 6(TRAF6), and downstream phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK), and phosphorylated nuclear factor-κB p65(p-NF-κB p65). Apoptosis of lung tissue cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL). The results showed that, compared with the control group, the model group exhibited significant pathological damage in lung tissue. The mRNA expression of IL-6, IL-1β, TNF-α, and CXCL6, as well as the protein levels of IL-17A, Act1, TRAF6, p-p38 MAPK/p38 MAPK, and p-NF-κB p65/NF-κB p65, were significantly increased, and the number of apoptotic cells was notably higher, indicating successful model establishment. Compared with the model group, both low-and high-dose groups of Bufei Decoction showed reduced pathological damage in lung tissue. The mRNA expression levels of IL-6, IL-1β, TNF-α, and CXCL6, and the protein levels of IL-17A, Act1, TRAF6, p-p38 MAPK/p38 MAPK, and p-NF-κB p65/NF-κB p65, were significantly decreased, with a significant reduction in apoptotic cells in the high-dose group. In conclusion, Bufei Decoction can effectively improve lung tissue damage and reduce inflammation in rats with K. pneumoniae. The mechanism may involve the regulation of the IL-17 signaling pathway and the reduction of apoptosis.
Animals ; Interleukin-17/metabolism* ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; Rats ; Male ; Klebsiella pneumoniae/physiology* ; Klebsiella Infections/immunology* ; Humans ; Lung/drug effects*

OBJECTIVE: This study aimed to evaluate the association between susceptibility genes and non-alcoholic fatty liver disease (NAFLD) in children with obesity. METHODS: We conducted a two-step case-control study. Ninety-three participants were subjected to whole-exome sequencing (exploratory set). Differential genes identified in the small sample were validated in 1,022 participants using multiplex polymerase chain reaction and high-throughput sequencing (validation set). RESULTS: In the exploratory set, 14 genes from the NAFLD-associated pathways were identified. In the validation set, after adjusting for sex, age, and body mass index, ECI2 rs2326408 (dominant model: OR = 1.33, 95% CI: 1.02-1.72; additive model: OR = 1.22, 95% CI: 1.01-1.47), C6orf201 rs659305 (dominant model: OR = 1.30, 95% CI: 1.01-1.69; additive model: OR = 1.21, 95% CI: 1.00-1.45), CALML5 rs10904516 (pre-ad dominant model: OR = 1.36, 95% CI: 1.01-1.83; adjusted dominant model: OR = 1.40, 95% CI: 1.03-1.91; and pre-ad additive model: OR = 1.26, 95% CI: 1.04-1.66) polymorphisms were significantly associated with NAFLD in children with obesity ( P < 0.05). Interaction analysis revealed that the gene-gene interaction model of CALML5 rs10904516, COX11 rs17209882, and SCD5 rs3733228 was optional ( P < 0.05), demonstrating a negative interaction between the three genes. CONCLUSION In the Chinese population, the CALML5 rs10904516, C6orf201 rs659305, and ECI2 rs2326408 variants could be genetic markers for NAFLD susceptibility.
Humans ; Non-alcoholic Fatty Liver Disease/genetics* ; Child ; Male ; Female ; Genetic Predisposition to Disease ; Case-Control Studies ; Exome Sequencing ; Adolescent ; Polymorphism, Single Nucleotide ; Obesity/complications* ; Pediatric Obesity/complications* ; China

OBJECTIVE: Poxviruses are zoonotic pathogens that infect humans, mammals, vertebrates, and arthropods. However, the specific role of ticks in transmission and evolution of these viruses remains unclear. METHODS: Transcriptomic and metatranscriptomic raw data from 329 sampling pools of seven tick species across five continents were mined to assess the diversity and abundance of poxviruses. Chordopoxviral sequences were assembled and subjected to phylogenetic analysis to trace the origins of the unblasted fragments within these sequences. RESULTS: Fifty-eight poxvirus species, representing two subfamilies and 20 genera, were identified, with 212 poxviral sequences assembled. A substantial proportion of AT-rich fragments were detected in the assembled poxviral genomes. These genomic sequences contained fragments originating from rodents, archaea, and arthropods. CONCLUSION Our findings indicate that ticks play a significant role in the transmission and evolution of poxviruses. These viruses demonstrate the capacity to modulate virulence and adaptability through horizontal gene transfer, gene recombination, and gene mutations, thereby promoting co-existence and co-evolution with their hosts. This study advances understanding of the ecological dynamics of poxvirus transmission and evolution and highlights the potential role of ticks as vectors and vessels in these processes.
Animals ; Poxviridae/physiology* ; Ticks/virology* ; Phylogeny ; Transcriptome ; Evolution, Molecular ; Poxviridae Infections/virology* ; Genome, Viral

ObjectiveTo analyze the evolutionary characteristics and genetic variations of the HA (hemagglutinin) and NA (neuraminidase) genes of influenza A(H1N1) viruses in Shanghai during 2024, to investigate their transmission patterns, and to evaluate their potential impact on vaccine effectiveness. MethodsFrom January to October 2024, throat swab specimens were collected from influenza like illness (ILI) patients at 4 hospitals in Shanghai. Real-time fluorescence ploymerase chain reaction (RT-PCR) was used for virus detection and isolation of H1N1 influenza viruses. Forty influenza A(H1N1) virus strains were sequenced using Illumina NovaSeq 6000 platform, followed by phylogenetic analyses, genetic distance analysis, and amino acid variation analyses of HA and NA genes. ResultsPhylogenetic tree of the HA and NA genes revealed that the 40 influenza A(H1N1) virus strains circulating in Shanghai in 2024 exhibited no significant geographic clustering, with a broad origin of strains and complex transmission chains. Genetic distance analyses demonstrated that the average intra-group genetic distances of HA and NA genes among the Shanghai strains were 0.005 1±0.000 6 and 0.004 6±0.000 6, respectively, which were comparable to or higher than those observed in global surveillance strains. Both HA and NA genes displayed frequent mutations. Compared to the 2023‒2024 and 2024‒2025 Northern Hemisphere A(H1N1) vaccine strains (WHO-recommended), the HA proteins of 40 Shanghai strains exhibited amino acid substitutions at positions 120, 137, 142, 169, 216, 223, 260, 277, 356 and 451, with critical mutations at positions 137 and 142 located within the Ca2 antigenic determinant. Furthermore, mutations in the NA protein were observed at positions 13, 50, 200, 257, 264, 339 and 382. ConclusionThe genetic background of the 2024 Shanghai influenza A(H1N1) virus strains is complex and diverse, and antigenic variation may affect vaccine effectiveness. Therefore, it is recommended to enhance genomic surveillance of influenza viruses, evaluate vaccine suitability, and implement more targeted prevention and control strategies against imported influenza viruses.

Objective: To develop a rapid quantification method for neutrophil extracellular traps (NETs) by quantifying neutrophils forming NETs (NETotic cells) on routine peripheral blood smears, and to evaluate the performance of an early warning model combining NETs with traditional coagulation indicators for risk of sepsis-induced coagulopathy (SIC) in septic patients. Methods: This prospective observational study was conducted in Wuxi People's Hospital Affiliated with Nanjing Medical University between May 2023 and May 2025. A total of 147 patients with sepsis (diagnosed based on Sepsis-3.0 criteria) who had not developed SIC [the international society on thrombosis and haemostasis (ISTH SIC score <4)] were enrolled. Blood samples were collected within 2 hours of admission. Neutrophil smudge cells (NETs%) were counted using an automated cell morphology analyzer. Serum levels of myeloperoxidase-DNA (MPO-DNA) complexes, circulating free DNA (cf-DNA) and sequential organ failure assessment (SOFA) scores were also determined. Based on progression to SIC (ISTH score ≥4) within 72 hours of admission, patients were categorized into a sepsis without SIC group (n=85) and a sepsis with SIC group (n=62). Risk factors were analyzed using binary logistic regression, receiver operating characteristic (ROC) curves were plotted, and the predictive value of NETs%, SOFA, and AT-Ⅲ for coagulation dysfunction was assessed. Results: The NETs% level was significantly higher in the sepsis with SIC group [8.50% (7.00, 11.50)] compared to both the healthy control group [1.00% (0.00, 2.00)] and sepsis without SIC group [4.40%(3.50, 6.50)] (P<0.01). NETs% was identified as an independent risk factor for SIC in sepsis patients. ROC analysis showed that the area under the curve (AUC) for predicting SIC was 0.90 for NETs%, 0.85 for MPO-DNA, and 0.79 for cf-DNA. The combined model of NETs% and SOFA score demonstrated the best performance, with an optimal cut-off value of 0.33, an AUC of 0.92, a sensitivity of 77%, and a specificity of 93%. Conclusion: NETs% shows promise as a novel biomarker for SIC. Peripheral blood smear morphology provides a simple, rapid, and cost-effective method for quantifying NETs%. NETs% enhances the early clinical identification of patients at high risk for SIC, and its combination with the SOFA score facilitates SIC prediction, offering a critical time window for initiating timely preventive interventions.

Objective To develop and validate a prediction model for risk of amputation in patients with diabetic foot ulcers(DFU)based on systematic review and meta-analysis.Methods The studies on the risk factors of amputation in DFU patients was retrieved by using subject words+free words.After screening,37 cohort studies were finally included,and the Newcastle-Ottawa scale(NOS)was used for quality evaluation.Meta-analysis was performed on the risk factors of amputation in DFU.Then a prediction model for DFU amputation risk were constructed based on the statistically significant risk factors in the meta-analysis.The corresponding β value was calculated based on the combined odds ratio(OR)value of each risk factor,and each risk factor was scored to establish a scoring system model.The clinical data of 453 DFU patients hospitalized in our department from 2021 to 2023 were collected as a validation cohort.Receiver operating characteristic(ROC)curve analysis was used to evaluate the model performance.The area under the curve(AUC)was calculated,and the optimal cutoff score was determined by calculation of the maximum Youden index through sensitivity and specificity.Results Our meta-analysis showed a cumulative amputation rate of approximately 34.65%in 11 779 DFU patients.The final risk prediction models include gangrene[OR=11.92(5.86～24.24)],ulcer depth[OR=4.93(2.52～9.64)],osteomyelitis[OR=3.19(2.36～4.29)],previous amputation history[OR=3.19(2.00～5.09)]and lower extremity arterial disease[OR=3.10(2.31～4.17)].According to the weights of each risk factor,the total score of the model is 76,and the optimal cut-off score is 36.5.The prediction model performed well,with an AUC value of 0.864(0.824,0.903),a sensitivity of 0.743,a specificity of 0.859,and an accuracy rate of 83.00%.Conclusion A prediction model for DFU amputation risk is developed based on risk factor scoring,and has good discrimination and calibration,providing effective scientific basis for clinical research and clinical decision-making related to DFU amputation.

Objective:To investigate the moyamoya disease (MMD) -related mitochondrial genes and to elucidate potential regulatory mechanisms.Methods:The GSE157628 and GSE189993 datasets of MMD from the Gene Expression Omnibus (GEO) database were downloaded as the training set and validation set, respectively. Mitochondrial-related gene sets were obtained from the MitoCarta 3.0 database. Weighted gene co-expression network analysis was used to screen the pathogenic module genes of MMD. Venn analysis was used to identify differentially expressed genes related to mitochondria. Functional enrichment analysis was used to elucidate potential molecular functions. Mitochondrial genes were screened using three machine learning methods, receiver operator characteristic (ROC) curve, and validation set expression analysis. A diagnostic nomogram model of MMD related features was developed based on diagnostic genes. The correlation between the identified genes and MMD immune infiltration characteristics, the biological pathways involved, and potential therapeutic drugs with interactions were analyzed.Results:A total of 15 differentially expressed mitochondria-related genes were identified based on GSE157628 dataset, involving the processes related to small molecule metabolism and mitochondrial function. Three machine learning algorithms, ROC curves and expression analysis finally identified two mitochondria-related diagnostic genes: NUDT8 and RDH13. The diagnostic nomogram model developed based on these two genes had good diagnostic ability for MMD. Immune infiltration analysis showed that there was significant difference in plasma cells between the MMD group and the control group. In addition, the two diagnostic genes interacted with several therapeutic drugs. Conclusion:NUDT8 and RDH13 are identified as potential mitochondrial-related genes of MMD, and they may serve as therapeutic targets for progressive vascular occlusion in MMD.

Objective:To investigate the effects of metformin on esophageal squamous cell carcinoma cell growth, migration and angiogenesis by regulating the expression of ALKBH3.Methods:Human esophageal cancer TE-1 cells were treated with different concentrations (0, 0.5, 1.0, 2.0, 4.0, 8.0 mmol/L) of metformin, and they were divided into a blank control group, low- (0.5 mmol/L), medium- (1.0 mmol/L), and high- (2.0 mmol/L) concentration metformin groups, a metformin (2.0 mmol/L) +pcDNA-NC group, and a metformin (2.0 mmol/L) +pcDNA-ALKBH3 group. The cell viability was determined by the CCK-8 method. The cell proliferation ability was detected by the clone formation assay. The cell migration and invasion abilities were examined by the Transwell assay. The cell apoptosis was detected by flow cytometry. The tube formation ability of cells was detected by the angiogenesis assay. A xenograft tumor model was constructed using 4- to 6-week-old male BALB/c thymus-less nude mice, which were divided into a model control group, a metformin group, a metformin+pcDNA-NC group, and a metformin+pcDNA-ALKBH3 group using a random number table method, and with six in each group. And the volume and weight of the tumor were measured. The protein expression levels of apoptosis-related proteins Bcl-2, Bax, ALKBH3 and vascular endothelial growth factor A (VEGF-A) were detected by Western blotting. The expression of CD31 protein was detected by immunohistochemistry.Results:After treating TE-1 cells with 0, 0.5, 1.0, 2.0, 4.0, and 8.0 mmol/L metformin for 48 hours, the cell viability was (100.00±0.00) %, (90.31±5.23) %, (81.25±8.65) %, (63.52±6.80) %, (54.64±5.35) %, and (31.48±4.21) %, respectively, with a statistically significant difference ( F=98.11, P<0.001). There were statistically significant differences in cell viability between 0.5, 1.0, 2.0, 4.0, 8.0 mmol/L and 0 mmol/L (all P<0.05). The IC 50 of metformin for TE-1 cells was 4.46 mmol/L. The numbers of colony formations of TE-1 cells in the blank control group, low-, medium-, and high-concentration metformin groups, metformin+pcDNA-NC group, and metformin+pcDNA-ALKBH3 group were 153.15±13.55, 134.80±11.62, 116.24±10.43, 93.17±8.85, 89.39±8.46, 110.26±7.21, respectively, with a statistically significant difference ( F=34.28, P<0.001); the numbers of colony formations of TE-1 cells in the metformin groups at different concentrations decreased significantly with the increase in metformin concentration (both P<0.05); compared with the metformin+pcDNA-NC group, the number of colony formations of cells in the metformin+pcDNA-ALKBH3 group increased ( P<0.05). The numbers of migration of TE-1 cells of 6 groups were 152.13±13.40, 133.85±10.72, 115.28±8.64, 91.16±7.89, 85.39±7.23, 116.85±8.36, the numbers of invasion were 135.22±10.77, 112.07±9.53, 86.30±7.45, 69.53±6.74, 65.81±5.65, 79.80±6.32, respectively, with statistically significant differences ( F=41.35, P<0.001; F=69.06, P<0.001); the numbers of migrated and invaded cells in the metformin groups at different concentrations decreased significantly with the increase in metformin concentration (all P<0.05); compared with the metformin+pcDNA-NC group, the numbers of migrated and invaded cells in the metformin+pcDNA-ALKBH3 group increased significantly (both P<0.05). The apoptosis rates of TE-1 cells in 6 groups were (3.22±1.13) %, (13.82±1.90) %, (22.67±2.53) %, (29.18±3.24) %, (26.84±2.75) %, and (16.36±1.63) %, respectively, with a statistically significant difference ( F=103.66, P<0.001); the apoptosis rates of cells in the metformin groups at different concentrations gradually increased with the increase in metformin concentration (both P<0.05); compared with the metformin+pcDNA-NC group, the apoptosis rate of cells in the metformin+pcDNA-ALKBH3 group was relatively lower ( P<0.05). The tubular structure of cells in blank control group was intact, and there were different degrees of damage to the tubular structure of cells in the low-, medium-, high- concentration metformin groups, the degree of damage to the tubular structure of cells in the metformin+pcDNA-ALKBH3 group was reduced. The numbers of cellular tubular structures of TE-1 cells in the 6 groups were 38.35±3.20, 27.15±2.64, 15.92±3.14, 7.39±1.50, 8.61±1.37, and 29.33±4.20, respectively, with a statistically significant difference ( F=113.92, P<0.001); the number of cellular tubular structures in the low-, medium-, and high- concentration metformin groups gradually decreased (both P<0.05); the number of cellular tubular structures in the metformin+pcDNA-ALKBH3 group was more than that in the metformin+pcDNA-NC group ( P<0.05). There were statistically significant differences in the protein expressions of Bcl-2, Bax, ALKBH3, and VEGF-A in TE-1 cells among 6 groups ( F=56.36, P<0.001; F=57.26, P<0.001; F=159.30, P<0.001; F=132.89, P<0.001); compared with the blank control group, the protein expressions of Bcl-2, ALKBH3, and VEGF-A in the metformin groups at different concentrations decreased, while the protein expression of Bax increased (all P<0.05); compared with the metformin+pcDNA-NC group, the protein expressions of Bcl-2, ALKBH3, and VEGF-A in the metformin+pcDNA-ALKBH3 group increased, and the expression level of Bax decreased (all P<0.05). The weights of tumors in the model control group, metformin group, metformin+pcDNA-NC group, and metformin+pcDNA-ALKBH3 group were (1.16±0.12), (0.46±0.05), (0.50±0.06), (1.19±0.14) g, the volumes of tumors were (878.36±108.93), (413.59±50.23), (439.78±51.39), (793.75±96.98) mm 3, with statistically significant differences ( F=96.61, P<0.001; F=51.90, P<0.001); the weight of tumors were lower and the volume of tumors were smaller in the metformin group than those in the model control group (both P<0.05), the weight of tumors were higher and the volume of tumors were bigger in the metformin+pcDNA-ALKBH3 group than those in the metformin group and the metformin+pcDNA-NC group (all P<0.05). CD31 was mainly distributed in the cytoplasm and cell membrane of tumor cells. There were statistically significant differences in the positive rates of CD31 and the protein expression levels of VEGF-A in transplanted tumor tissues among 4 groups ( F=7.12, P=0.002; F=48.81, P<0.001); the positive rate of CD31 and the protein expression level of VEGF-A in the metformin group were lower than those in the model control group; the positive rate of CD31 and the protein expression level of VEGF-A in the metformin+pcDNA-ALKBH3 group were higher than those in the metformin group and the metformin+pcDNA-NC group (all P<0.05) . Conclusions:Metformin may inhibit the proliferation, migration, and tumor angiogenesis of esophageal squamous cell carcinoma by reducing ALKBH3 expression.

Objective To evaluate the efficacy and safety of different Chinese medicine injections combined with angiotensin converting enzyme inhibitor/angiotensin Ⅱ receptor blocker(ACEI/ARB)in the treatment of proteinuria in diabetic nephropathy by using network Meta-analysis.Methods Clinical randomized controlled trials of Chinese medicine injection combined with ACEI/ARB for the treatment of proteinuria in early diabetic nephropathy were retrieved from the domestic databases such as China National Knowledge Infrastructure(CNKI),China Science and Technology Journal Database(VIP),Wanfang Data Knowledge Service Platform(Wanfang Data),and China Biology Medicine Disc(CBM),as well as from the oversea databases such as PubMed,EMbase,Web of Science,and Cochrane Library from database inception to October 2023.The Cochrane risk of bias tool ROB2.0 was used to evaluate the quality of the included literature,and traditional Meta-analysis and network Meta-analysis were performed by using R4.3.2 and Stata18.0.Results A total of 72 trials were included,involving 13 Chinese medicine injections,namely Huangqi Injection,Xuesaitong Injection,Puerarin Injection,Denzhan Xixin Injection,Breviscapin Injection,Danhong Injection,Xueshuantong Injection,Yinxingye Injection,Shuxuetong Injection,Yinxing Damo Injection,Danshen Injection,Shenkang Injection,and Danshen Ligustrazine Injection.The primary outcomes were urinary albumin excretion rate(UAER),24-hour urinary total protein(24hUTP),serum creatinine(SCr),and urine albumin-to-creatinine ratio(UACR).The results of traditional Meta-analysis showed that Chinese medicine injection combined with ACEI/ARB was more effective on reducing the levels of UAER[MD=-26.14,95%CI(-29.39,-22.89)],24hUTP[(MD=-0.22,95%CI(-0.27,-0.18)],SCr[MD=-0.34,95%CI(-0.50,-0.19)],and UACR[MD=-9.40,95%CI(-15.64,-3.16)]than ACEI/ARB alone,and the differences were all statistically significant(P<0.05).The results of network meta-analysis showed that the efficacy of Dengzhan Xixin Injection combined with ACEI/ARB was the optimum in terms of reducing UAER,Danshen Ligustrazine Injection combined with ACEI/ARB was the optimum in terms of decreasing 24hUTP,Shenkang Injection combined with ACEI/ARB was the optimum in terms of decreasing SCr,and Danhong Injection combined with ACEI/ARB had the best efficacy on improving UACR.Conclusion The efficacy and safety of Chinese medicine injection combined with ACEI/ARB in the treatment of proteinuria in early diabetic nephropathy are superior to those of ACEI/ARB alone in terms of lowering UAER,24hUTP,SCr,and UACR.However,due to the limitations of the quality and number of included literatures,the conclusions still need to be confirmed by more high-quality randomized controlled trials.